Skeletal calcification patterns in the sea urchin Tripneustes gratilla elatensis (Echinoidea: Regularia)

A comprehensive study on skeletal calcification in the regular echinoid Tripneustes gratilla elatensis was carried out between 1979 and 1981 in the northern Gulf of Eilat (Red Sea), employing size measurement, allometry and radiotracer techniques. Uptake rates of the isotope ⁴⁵Ca were used to estimate the calcification rate of whole tests. Calcification rates of the different skeletal parts and the ⁴⁵Ca uptake along the sutural margins of individual plates were also measured. Whole-test calcification rates for juvenile individuals, 11 to 13 mm in diameter, relative to their skeletal Ca dry weight, calculated using the allometric relationship between skeletal dry weight and diameter, were 1.5 to 1.8% d^-1 for 4 to 8 h of incubation, while calcification rates obtained from periodical size measurements and allometrical constants amounted to 2.75% d^-1. The apparent discrepancy between these results is explained partly by the short duration of the incubations, during which the internal calcium pools, apparently in the coelom, were not fully saturated with the radioactive tracer. This discrepancy decreased when longer radioactive incubations were used, or when a post-incubational deposition of ⁴⁵Ca from these pools was allowed. The size of these pools is roughly 10 mol Ca per 13-mm individual, and it resides probably in the coelom where Ca concentration is 3 times higher than in seawater. The calcification rates given as percentage of skeleton added per day, both for the radioactive and allometric methods, decreased with the urchins' size. The calcification patterns of various components of the skeleton generally fitted their allometrical trends — a relative decrease in the size of the spines, apical plates and lantern. The teeth calcify rapidly to compensate for the constant erosion of their tips. Calcification per plate decreased exponentially from the apex to the peristome. Plate-edge calcification patterns of both interambulacral and ambulacral plates fitted patterns derived from size increment measurements, using natural growth-lines. Typically, most plates grow more horizontally (latitudinally) than vertically. It is suggested that the vertical vs horizontal calcification ratio (v/h) determines the individual plate growth and affects the whole test morphology.     

Mechanisms for the uptake of inorganic carbon by two species of symbiont-bearing foraminifera

The mechanisms for uptake of inorganic carbon (Ci) for photosynthesis and calcification of a perforate foraminifer, Amphistegina lobifera Larsen, and an imperforate species, Amphisorus hemprichii Ehrenberg, from the Gulf of Eilat, Red Sea were studied in 1986–1987 using ¹⁴C tracer techniques. Total Ci uptake of A. lobifera and photosynthetic carbon uptake of A. hemprichii fit the Hill-Whittingham equation that describes the overall rate of enzymatic reactions that are provided with their substrate through a diffusion barrier. This suggests that diffusion is the rate limiting step for total Ci uptake in A. lobifera. Photosynthesis by the isolated symbionts and uptake of CO₃ ^2- for calcification obey Michaelis-Menten kinetics indicating that enzymatic reactions determine the rate of the separate processes. Both photosynthesis and calcification can be inhibited without affecting each other. Calcification rates in A. lobifera were optimal at Ca levels around normal seawater concentration and were sensitive to inhibitors of respiratory adenosine triphosphate (ATP) generation and Ca-ATP-ase. This indicates that Ca uptake is also active. Calcification rates of A. hemprichii increased linearly as a function of external Ci concentration over the entire experimental range (0 to 4 mM Ci). In contrast, photosynthetic rates showed Hill-Whittingham type kinetics. The dependence of calcification on the CO₃ ^2- concentration was also linear, suggesting that its diffusion is the rate limiting step for calcification in A. hemprichii. Increasing Ca concentrations yielded higher calcification rates over the entire range measured (0 to 40 mM Ca). Calcification in A. hemprichii was less sensitive to inhibitors of ATP generation than in A. lobifera, suggesting that in A. hemprichii energy supply is less important for this process.     

Calcium exchange and the measurement of calcification rates in the calcareous coralline red alga Amphiroa foliacea

⁴⁵Ca washout data of living thalli of Amphiroa foliacea Lamouroux in the light and dark show that there are three kinetically distinct Ca²⁺-exchanging compartments with approximate half-times (t 1/2) of 300, 20 and 3 min. The two slower compartments appear to be exchange from organic Ca²⁺-binding components of the cell wall, while the fast compartment probably represents exchange on the CaCO₃ crystal surface. Killed and decalcified thalli have a fourth compartment, with a t 1/2 of 20 to 35 min (other compartment half-times are 300, 40, 3 min), which has been identified as the greatly increased intercellular space produced during drying and decalcification. The ⁴⁵Ca and ¹⁴C uptake data show that a large proportion of the label initially taken up is into compartments other than the CaCO₃. As a result of this uptake, binding, and exchange of radioisotope, significant errors occur during the measurement of calcification rates, unless a kinetic analysis is carried out. Using such a technique, CaCO₃ calcification rates of A. folicea were measured with ⁴⁵Ca or ¹⁴C as tracers. Light stimulates calcification by up to 2.6 times, depending upon the age of the plant. Young segments have a markedly higher rate of calcification and photosythesis than do the older segments.     

Estimation of coral growth-rates from laboratory 45Ca-incorporation rates

Laboratory ⁴⁵Ca-incorporation rates in hermatypic coral skeletons have previously been used successfully as an index of physiological function. This laboratory method would become more meanigful if it also provided an absolute measure of coral growth rates. In two coral species, Porites compressa and Pocillopora damicornis, ⁴⁵Ca incorporation rates were obtained from short (0.5 h) laboratory incubations using apical (determined as fast growing) portions of freshly collected coral branches. ⁴⁵Ca exchange across the coenosarc was not significant and not corrected for, whereas diurnal fluctuation in ⁴⁵Ca in Pocillopora damicornis was significant and a necessary correction. A calculated surface area is used to express calcification rate. Typical growth rates calculated from the ⁴⁵Ca-incorporation rates were 20 and 6 mm/year for Porites compressa and Pocillopora damicornis, respectively. These rates are considerably higher than those previously obtained in the laboratory, and compare favorably with field growth rates — 24 and 14 mm/year, respectively.     

Coral calcification: Sources of error in radioisotope techniques

Isotopic exchange occurs between coral skeleton and ⁴⁵Ca⁺⁺ and H¹⁴CO ₃ ^- in seawater. Exchange of ¹⁴C onto skeletons is more rapid than exchange of ⁴⁵Ca⁺⁺. Exchange of ¹⁴C from skeletons to seawater takes place more slowly than exchange of ⁴⁵Ca⁺⁺ to seawater. When living coral is incubated in the dark with radioisotopes for 1 h, the tissues contain considerably more radioactivity than is associated with the skeleton. The tissue radioactivity reflects permeation of tissues and coelenteron by radioactive compounds from the incubation seawater. Addition of alkalis to cardioactive seawater results in a radioactive precipitate, part of which becomes associated with any coral skeleton present, and part of which forms on the wall of the containing vessel. Strong alkali removes biologically-deposited radioisotope from coral skeletons. Deposition, of ¹⁴C from H¹⁴CO ₃ ^- in skeletons of living coral incubated in the dark is greater than in dead coral. The reverse situation occurs with ⁴⁵Ca⁺⁺.     

Influence of some coral reef communities on the calcium carbonate budget of Tiahura reef (Moorea,French Polynesia)

The calcium carbonate budget of coral reefs is the result of the interaction of the processes of calcification and biological degradation, and is reflected in the chemical properties of the seawater overlying the reefs. A series of experiments at Moorea Island (French Polynesia) in 1988 monitored the diurnal and nocturnal variations in the chemical properties of seawater under field and laboratory conditions. Our results revealed that in the study area (Tiahura barrier reef flat), the calcium carbonate budget varied over space and time as a function of location in the water current. Two in-situ sites were investigated; one was situated 100 m from the algal crest of the barrier reef, the other 300 m further downstream. As a result of cumulative upstream events, the daily net calcification was ten times higher at the downstream (5.22 gm^-2 d^-1) than at the upstream (0.45 gm^-2 d^-1) site. The carbonate uptake by in situ Porites lobata in enclosures (8 kgm^-2 yr^-1) was ten times higher than the uptake by the whole community in the surrounding water (0.8 kgm^-2 yr^-1) and five times higher than that recorded for P. lobata in laboratory experiments (1.4 kgm^-2 yr^-1), where illumination levels were 10% of in situ levels. In laboratory experiments, the planktonic fraction of the seawater had no perceptible influence on the calcium carbonate budget. In the absence of bioeroders, living coral totally depleted the carbonate content of the seawater (3.7 gm^-2d^-1). Bioerosive organisms played an important role in restoring this calcium carbonate; e.g. sea urchins grazing on algal turf covering dead coral ingested CaCO₃ and released this as a carbonate powder (1.26 gm^-2d^-1); a form of carbonate which is extremely accessible to chemical dissolution.     

Coral mucus stable isotope composition and labeling: experimental evidence for mucus uptake by epizoic acoelomorph worms

Mucus released by scleractinian corals can act as an important energy and nutrient carrier in coral reef ecosystems, and a distinct isotopic signature would allow following the fate of this material. This study investigates the natural C and N stable isotopic signatures of mucus released by four scleractinian coral genera (Acropora, Fungia, Pocillopora and Stylophora) in comparison with those of suspended particulate organic matter (POM) in seawater of a Northern Red Sea fringing coral reef near Aqaba, Jordan. The natural δ¹³C and δ¹⁵N signatures of coral mucus differed significantly from seawater POM for the majority of seasonal comparisons, but were inappropriate for explicit tracing of mucus in the coral reef food web. Thus, a labeling technique using stable isotope tracers (¹³C and ¹⁵N) was developed that produced δ¹³C values of up to 122 ± 5‰ (mean ± SE) and δ¹⁵N of up to 2,100 ± 151‰ in mucus exuded by Fungia corals. ¹³C and ¹⁵N-enriched compounds were rapidly (within 3 h) and light-dependently transferred from the endosymbiotic zooxanthellae to the mucus-producing coral host. The traceability of ¹⁵N-labeled mucus was examined by evaluating its uptake and potential utilization by epizoic acoelomorph Waminoa worms naturally occurring on a range of scleractinian coral taxa. This tracer experiment resulted in uptake of coral mucus by the coral-associated acoelomorphs and further demonstrated the possibility to trace stable isotope-labeled coral mucus by revealing a new trophic pathway in coral reef ecosystems.     

Uptake of inorganic carbon and internal carbon cycling in symbiont-bearing benthonic foraminifera

Incorporation rates of inorganic carbon and its distribution between the organic matter and the skeleton have been measured using ¹⁴C tracer techniques on two species of symbiont-bearing benthonic foraminifera in the Gulf of Elat: Amphistegina lobifera (a perforate species) and Amphisorus hemprichii (an imperforate species). Under constant experimental conditions, incorporation rates of the radiotracer become linear with time after several hours in A. hemprichii and after one day in A. lobifera. A. lobifera showed a lag time of 24 h for skeletal incorporation, whereas in A. hemprichii uptake into the skeleton started within 2 h. Pulse-chase incubations in radioactive seawater, followed by unlabelled incubations, demonstrate transfer of photosynthetically acquired ¹⁴C into the skeleton of A. lobifera. No such transfer was found in A. hemprichii. The total ¹⁴C uptake by A. lobifera increased during the first 24 h of cold chase incubation. This increase suggests the existence of an internal inorganic carbon pool that was lost (probably evaporated) during the analysis of pulse incubations. However, during the following chase incubations, the ¹⁴C in this pool was incorporated mainly into the skeleton and retained during analysis, causing the increase in the total uptake. No such increase was found in A. hemprichii. Additional ¹⁴C uptake experiments on other species of the genera Operculina, Heterostegina and Borelis suggest that the differences in pathways for incorporation of carbon between A. lobifera and A. hemprichii can be generalized to the perforate and imperforate foraminiferal groups. In perforate species, respired carbon originally taken up through photosynthesis is partly recycled into the skeleton. In imperforate species such a transfer has not been demonstrated. Perforate species seem to have a large internal inorganic carbon pool which serves mainly for calcification and possibly also for photosynthesis, while imperforate species may take up carbon for calcification directly from seawater or have a very small inorganic carbon pool.     

Effect of temperature and temperature adaptation on calcification rate in the hermatypic coral Pocillopora damicornis

Using ⁴⁵Ca incorporation into the coral skeleton as a measure of calcification rate, the effect of temperature on clacification rate was studied in the hermatypic coral Pocillopora damicornis. Both immediate and long-term (adaptation) effects were investigated. Temperature has a marked effect on rate — an effect that varies depending on the temperature history of the coral (i.e., temperature adaptation occurs). P. damicornis showed both 27° and 31°C temperature optima, one or the other being dominant depending on the natural water temperature to which the coral was adapted. The two optimum temperatures may indicate two isoenzymes or two alternate metabolic pathways involved in the calcification process.     

Cohabitation of a coral reef sponge and a colonial scyphozoan

A sensitive experimental protocol using cloned corals (hereafter microcolonies) of the branching scleractinian coral Stylophora pistillata and ⁴⁵Ca has been developed to enable reproducible measurements of physiological and biochemical mechanisms involved in calcium transport and compartmentalization during coral calcification. Cloned S. pistillata microcolonies were propagated in the laboratory from small fragments of parent colonies collected in 1990 in the Gulf of Aqaba, Jordan. Cloned microcolonies have several intrinsic properties that help to reduce unwanted biological variability: (1) same genotype; (2) similar sizes and shapes; and (3) absence of macroscopic boring organisms. Errors specifically associated with long-standing problems to do with isotopic exchange were further reduced by producing microcolonies with no skeletal surfaces exposed to the radioisotope-labelled incubation medium. The value of the technique resides principally in its superior ability to elucidate transportation pathways and processes and not in its ability to quantitatively estimate calcium deposition by corals in nature. We describe here a rapidly exchangeable calcium pool in which up to 90% of the radioactive label taken up during incubations is located. This pool (72.9±1.4 nmol Ca mg^-1 protein) is presumably located within the coelenteric cavity as suggested by the following: (1) it has 4-min half-time saturation kinetics; (2) the accumulation of calcium is linearly correlated with the calcium concentration of sea-water; and (3) its insensitivity to metabolic and ion transport inhibitors indicate that membranes do not isolate this compartment. Washout of this large extracellular pool greatly improved estimates of calcium deposition as evidenced by 10 to 40% reduction in coefficients of variation when compared with previous ⁴⁵Ca²⁺ methods described in the literature. Comparisons of calcification measurements simultaneously carried out using the alkalinity anomaly technique and the ⁴⁵Ca protocol described here show that the correlation coefficient of both techniques is close to 1. Unlike previous reports, our ⁴⁵Ca²⁺-derived measurements are slightly lower than those computed from the alkalinity depletion technique.     

Bacterial colonization and endocytosis on the gill of a new limpet species from a hydrothermal vent

Calcification rates in different fragments along branches of the hermatypic coral Stylophora pistillata were tested in the laboratory using a new technique, the optic glassfiber method. By this method, the tested colony remains constantly in dark conditions while a narrow beam of light, transferred through the optic fiber, illuminates a small distinct point of coral tissue (on a branch tip or base). The selected illuminated portion of the branch serves as the experimental fragment, while all the other parts of the same colony serve as the dark controls. The results indicate that significantly more calcium is incorporated in the tip fragments than in the bases, both in light and in dark conditions (4.1 to 13.2 times more). Illumination of the tips or the bases did not stimulate or enhance the calcification rates of these fragments. Thus, in all colonies tested, the calcification rates of the illuminated fragments were not significantly different from the average rates of other similar, non-illuminated fragments of the same colony. It is suggested that light does not directly enhance calcification in hermatypic corals, but rather, that light enhances O₂ production, which consequently stimulates coral metabolism. Our preliminary results indicate that calcification rates recorded in aerated dark experiments are significantly higher than calcification rates of non-aerated dark controls.     

Short-term growth measurements of corals using an accurate buoyant weighing technique

An accurate method for determining the growth rates of the skeleton of isolated branch tips (nubbins) of corals over intervals of less than 24 h is described. The skeletal weight of the coral was estimated from its buoyant weight in seawater whose density had been accurately determined. The coral tissues accounted for between 1 and 5% of the total buoyant weight in Pocillopora verrucosa and Acropora humilis with differing relative tissue biomass. After correcting for tissue buoyant weight, predictions of skeletal weight were accurate to within 1%. The method was used to estimate the growth of sample nubbins of Porites porites of similar diameter, in 2 m of water at Discovery Bay, Jamaica. Since growth of these branch tips is apical, growth rate could be expressed without correction for the size. The mean 24 h skeletal growth rate ranged between 40 and 47 mg. Differences could be measured between day-time and night-time growth, the day: night ratio being 3.7. The method also showed that P. porites virtually ceases calcification during the 4 to 5 d periods that it becomes enclosed in a mucus tunic. Nubbins of P. porites attached to the reef at different locations showed clear differences in growth rate with depth, and between clear and turbid water sites. The growth rate of nubbins was compared with that of branch tips of whole corals by measuring the linear extension after staining with Alizarin Red S. After 3 1/2 mo, the mean linear extension was 4.1 mm in each case, indicating that the growth rate of nubbins is the same as that of branch tips of the whole colony. It is suggested that this buoyant weighing technique will find applications in laboratory experiments with calcification mechanisms and as a bioassay on reefs exposed to environmental stress.Contribution No. 464 of the Discovery Bay Marine Laboratory     

Silica uptake of the marine sponge Halichondria panicea in Kiel Bight

The silica uptake of Halichondria panicea Pallas, 1766 was measured in laboratory experiments from February until August 1995. Uptake rates were determined by measuring the decrease of dissolved silica in the surrounding seawater. All specimens were collected in Kiel Bight and maintained in aquaria up to several weeks prior to the experiments. Up to 5.27 μmol Si g⁻¹ AFDW h⁻¹ were incorporated by the sponges, with a strong, size-independent individual variation. A positive correlation between the content of dissolved silica in the seawater and the silica uptake was found. Temperature had no decisive effect on uptake rates. The nutritional condition of the sponges turned out to be of great importance. After 1 wk of starvation, Halichondria panicea uptake rates were only 15% of the previous amount, which indicates that silica transport is an energy-consuming process. During the most intense phase of reproduction activity in spring, female specimens showed a significant drop in their silica uptake. Obviously they did not produce spicules during this time. An attempt to estimate the influence of H. panicea on its habitat as a consumer of dissolved silica led to the conclusion that in summer the sponges might affect the phytoplankton species composition by competing with diatoms for dissolved silica. Received: 4 August 1996 / Accepted: 14 October 1996     

The synergistic effects of hydrogen peroxide and elevated seawater temperature on the metabolic activity of the coral <Emphasis Type="Italic">Galaxea fascicularis</Emphasis>

We examined quantitative changes in the metabolism of the coral Galaxea fascicularis caused by increases in both hydrogen peroxide (H₂O₂) concentration and seawater temperature. Seawater temperatures were maintained at 27 or 31°C in a well-controlled incubation chamber, and three levels of H₂O₂ concentration (0, 0.3, 3.0 μM) were used in experimental treatments. Gross primary production, calcification rates and respiration rates were all affected by increased H₂O₂ concentrations and high seawater temperatures. Individual treatments of high H₂O₂ or elevated seawater temperature alone caused significant declines in coral photosynthesis and calcification rates within the 3-day incubation period. The synergistic effect of high H₂O₂ combined with high seawater temperature resulted in a 134% increase in respiration rates, which surpassed the effect of either H₂O₂ or high seawater temperature alone. Our results suggest that both high H₂O₂ concentrations and elevated temperatures in seawater can strongly affect coral metabolism; however, these effects cannot be estimated by simply summing the effects of individual stress parameters.     

Activation energy for skeletal aragonite deposited by the hermatypic coral Platygyra spp.

Average radial growth rates of the hemispherical aragonite colonies deposited by the Indo-Pacific scleractinian reef coral Platygyra spp. were determined by measuring the thickness of density variations in the skeleton that are revealed by X-radiography. Ninety-one specimens from 21 localities were examined, but only 54 of these exhibited well-defined growth-banding. The apparent temperature dependence of growth rate is linear over the range 23.9° to 29.3°C, averaging 5.4±0.94 mm/year at 24°C, 8.0±0.42 mm/year at 27°C, and 9.7 ±0.58 mm/year at 29°C (90% confidence limits). Expression of the influence of temperature on growth rate in terms of the Arrhenius equation yields an activation energy of 20,680 cal/mole, which is comparable to values for typical biological reactions, but is only half that reported for skeletogenesis in another reef coral, Pocillopora damicornis, on the basis of controlled incubation studies involving ⁴⁵Ca uptake.     

Application of the 45Ca tracer method for determination of calcification rates in calcareous algae: Effect of calcium exchange and differential saturation of algal calcium pools

Calcium exchange and differential saturation of algal calcium pools complicate the application of the ⁴⁵Ca tracer method for the determination of net deposition rates of calcium in calcareous algae. The kinetics of ⁴⁵Ca uptake is critically reevaluated, and it is shown that incorporation occurs in two stages. A fast stage, saturating the exchangeable calcium pools; and a slow stage, giving rise to net deposition. The reliability of the method much depends on the determination of the second rate constant. Calcium net-deposition rates are obtained from the expression , where represents the second rate constant and S _SW the specific activity of the seawater. Calcium exchange and recycling of tracer would lower the second rate constant, hence the method will give rise to minimum values. The application of the method is demonstrated for the following algae: Halimeda incrassata, H. opuntia, Penicillus pyriformis, Udotea flabellum, Cymopolia barbata, Padina sanctae crucis and Amphiroa fragilissima; the results show close agreement of data with independent chemical estimates.Sonderforschungsbereich 95 der Universität Kiel, Publication No. 200.Contribution No. 732 from the Bermuda Biological Station for Research.     

Calcification in the staghorn coral Acropora acuminata: Variations in apparent skeletal incorporation of radioisotopes due to different methods of processing

Pieces of branch from the staghorn coral Acropora acuminata were incubated with ⁴⁵CaCl₂ and NaH¹⁴CO₃ under identical conditions in the light or in the dark. Specimens were then processed in different ways. All specimens were placed in N KOH to digest tissues. Some were placed in KOH immediately after incubation; others were placed in KOH after 2 h washing, or after 2 h extraction with methanol-chloroformwater. Specimens were washed in running fresh water or running seawater; some were killed in liquid N₂ before washing. Radioactivity associated with skeleton and tissues was determined. The method of processing profoundly affected the results. In dark incubations, there was up to a four-fold difference in apparent skeletal incorporation of ⁴⁵Ca⁺⁺ between average values obtained for the different treatments. For ¹⁴C incorporation, there was a difference of up to 2.5 times. In light incubations, skeletal incorporation of both radioisotopes showed a two-fold difference between high and low average values obtained for the different treatments.     

Determination of ammonium uptake and regeneration rates using the seawater dilution method

We have developed a method for the determination of ammonium uptake and regeneration rates applying the principle of the seawater dilution technique. The method is based on the separation of uptake and regeneration processes in the dilution series. A model is used to estimate ammonium uptake and regeneration rates simultaneously, in addition to phytoplankton growth and grazing rates. The method was applied to dilution experiments conducted during a two-year study of the upwelling region off Oregon, USA. Ammonium uptake and regeneration rates determined with our method ranged from 0.5 to 3 mol l^-1d^-1 and from 0.2 to 2.9 mol l^-1d^-1, respectively. These values agree well with those from other studies applying ¹⁵N tracer techniques in the same or similar environments. We found a close coupling between ammonium uptake and regeneration, and a strong relationship between ammonium regeneration and grazing rates. In addition, the nutritional status of the phytoplankton community could be assessed by comparing instantaneous ammonium uptake rates with the specific phytoplankton growth rates. Using the dilution technique to determine ammonium uptake and regeneration rates of the plankton community is a promising alternative to the application of tracer techniques conventionally used to determine these rates.     

Experimental ecology of the temperate scleractinian coral Astrangia danae

The combined effects of temperature, light and symbiont density on the metabolic rate and calcification of the temperate coral Astrangia danae were studied experimentally using colonies containing different concentrations of zooxanthellae. After acclimation to five temperatures between 6.5° and 27°C, and incubation at three light levels and in darkness, respiration and photosynthesis were measured and corrected for rates due to commensals alone. Calcification rates were regressed on zooxanthellae concentration and production in order to define “symbiotic” and “non-symbiotic” averages, and the enhancement of calcification by symbiotic interactions in the polyps. Respiration by the polyparium varied less with temperature between 11.5° and 23°C than that of the commensals, suggesting physiological acclimation by the coral tissue. In-vivo zooxanthellae photosynthesis increased linearly with temperature and was near its maximum at 400 μEin m^?2 s^?1, but the photosynthesis of the endolithic algae of the corallum varied little between 11.5° and 27°C. Calcification at any given temperature was near its maximum at 40 μEin m^?2 s^?1 in both symbiotic and non-symbiotic corals. CaCO₃ deposition increased linearly with temperature in non-symbiotic colonies and in symbiotic colonies incubated in the dark. In symbiotic colonies, calcification in the light increased above these basic rates as temperature rose above 15°C. Below 15°C, symbiotic interactions failed to stimulate calcification, apparently due both to a lowering of zooxanthellae photosynthesis and to a decrease in the enhancing effect of any given level of primary production.